CLONING AND CHARACTERIZATION OF RABBIT PANCREATIC COLIPASE

1. Among the digestive enzymes synthesized by pancreas. lipase is the principle lipolytic enzyme which hydrolyses dietary glycerides. 2. For its action it requires a coenzyme, colipase. 3. The molecular mechanisms of the interaction of these two are not fully understood. 4. Further, molecular events that regulate and influence lipid absorption are ill defined. 5. The rabbit is the conventional animal model for the study of lipid absorption. We have undertaken the molecular cloning, and characterization of rabbit pancreatic colipase, the coenzyme for pancreatic lipase. 6. Colipase has been cloned from a gt 11 library of an adult rabbit pancreatic cDNA by probing with an oligonucleotide derived from human colipase sequence. 7. The total reading frame consists of 321 nucleotides coding for 90 amino acids of the functional protein and 17 nucleotides of the leader peptide. 8. Northern blot analysis revealed a distinct band around 0.5 kb. Comparison with other species revealed an over all homology of 75% at the nucleotide level. 9. At the amino acid level highest conservation is observed at the lipase-binding region (AA 53-73). 10. Rabbit enzyme also retained the N-terminal pentapeptide of it preform. 11. The regions of homology and conservation may aid to define the sites of interaction of colipase with lipase.